Transgenic plants expressing MinD or MinE and an efficient method for plant chloroplast transformation and gene expression

Abstract

The present invention concerns transgenic plant which contain large chloroplasts. The transgenic plant of the present invention comprises within its genome a foreign MinD or MinE gene or a foreign gene which expresses a protein which has the same functional activity as the Arabidopsis thaliana MinD or MinE protein. The present invention further concerns a method of producing the transgenic plants of the present invention which contain large chloroplasts. Finally, the present invention concerns a method of transforming the chloroplasts genome of the transgenic plants of the present invention which contain large chloroplasts.

We claim:  
     
         1 . A vector comprising an exogenous gene which encodes a protein which has the same functional activity as a protein encoded by the  Arabidopsis thaliana  MinE or MinD gene and which when expressed in a plant cell causes the plant cell to have enlarged and/or a reduced number of chloroplasts.  
     
     
         2 . A cell comprising the vector of  claim 1 .  
     
     
         3 . A tissue culture comprising cells of  claim 2 .  
     
     
         4 . A seed comprising the vector of  claim 1 .  
     
     
         5 . The vector according to  claim 1 , wherein said exogenous gene is derived from  Arabidopsis thaliana.    
     
     
         6 . The vector according to  claim 1 , wherein the exogenous gene is an exogenous MinD gene.  
     
     
         7 . The vector according to  claim 6 , wherein said exogenous MinD gene is derived from  Arabidopsis thaliana  MinD gene.  
     
     
         8 . The vector according to  claim 1 , wherein the exogenous gene is an exogenous MinE gene.  
     
     
         9 . The vector according to  claim 8 , wherein said exogenous MinE gene is derived from  Arabidopsis thaliana  MinE gene.  
     
     
         10 . A transgenic plant comprising within its nuclear genome an exogenous gene, wherein said exogenous gene encodes a protein which has the same functional activity as a protein encoded by the  Arabidopsis thaliana  MinE or MinD gene and which when expressed in a plant cell causes the plant cell to have enlarged and/or a reduced number of chloroplasts.  
     
     
         11 . The transgenic plant according to  claim 10 , wherein said exogenous gene is derived from  Arabidopsis thaliana.    
     
     
         12 . The transgenic plant according to  claim 10 , wherein said exogenous gene is an exogenous MinD gene.  
     
     
         13 . The transgenic plant according to  claim 12 , wherein said exogenous MinD gene is derived from  Arabidopsis thaliana  MinD gene.  
     
     
         14 . The transgenic plant according to  claim 10 , wherein said transgenic plant is a tobacco plant.  
     
     
         15 . The transgenic plant according to  claim 10 , wherein said exogenous gene is an exogenous MinE gene.  
     
     
         16 . The transgenic plant according to  claim 15 , wherein said exogenous MinE gene is derived from  Arabidopsis thaliana  MinE gene.  
     
     
         17 . A method of transforming the chloroplast genome of a plant, said method comprising the steps of: 
 A) producing a nuclear transgenic plants which contains large chloroplasts by: 
 i) providing a vector comprising an exogenous gene which encodes a protein which has the same functional activity as a protein encoded by the  Arabidopsis thaliana  MinE or MinD gene and which when expressed in a plant cell causes the plant cell to have enlarged and/or a reduced number of chloroplasts; and,  
 ii) transforming the nuclear genome of a plant with said vector which comprises said exogenous gene; and,  
   B) transforming the chloroplast genome of said nuclear transgenic plant with a vector which comprises Gene of interest.    
     
     
         18 . The method of  claim 17 , wherein said exogenous gene is derived from  Arabidopsis thaliana.    
     
     
         19 . The method of  claim 17 , wherein said exogenous gene is an exogenous MinD gene.  
     
     
         20 . The method of  claim 19 , wherein said exogenous MinD gene is derived from  Arabidopsis thaliana  MinD gene.  
     
     
         21 . The method of  claim 17 , wherein said nuclear transgenic plant is a tobacco plant.  
     
     
         22 . A chloroplast transgenic plant produced by the method of  claim 17 .  
     
     
         23 . The method of  claim 17 , wherein said exogenous gene is an exogenous MinE gene.  
     
     
         24 . The method of  claim 23 , wherein said exogenous MinE gene is derived from  Arabidopsis thaliana  MinE gene.  
     
     
         25 . A method of selecting for plants that are chloroplast transgenics but not nuclear transgenics, wherein said method comprises: 
 A) crossing a plant produced by the method of  claim 22  with a wild-type plant; and,    B) segregating out the plants which express the exogenous gene or genes of interest in the chloroplast genome and further do not express the exogenous gene in the nuclear genome by identifying which plants have normal chloroplast size and number and have the desired characteristic produced by the exogenous gene expressed in the chloroplast genome.    
     
     
         26 . The method of  claim 25 , wherein said exogenous gene is derived from  Arabidopsis thaliana.    
     
     
         27 . The method of  claim 25 , wherein said plant which is a chloroplast transgenic is a tobacco plant.  
     
     
         28 . A method of producing a transgenic plant which comprises one or a few large chloroplasts, said method comprising the steps of: 
 A) producing a vector comprising an exogenous gene which encodes a protein which has the same functional activity as a protein encoded by the  Arabidopsis thaliana  MinE or MinD gene and which effects a plant cell by allowing for the expression of only one or a few large chloroplasts; and    B) transforming the nuclear genome of a plant with said vector.    
     
     
         29 . The vector according to  claim 28 , wherein said exogenous gene is derived from  Arabidopsis thaliana.    
     
     
         30 . The vector according to  claim 28 , wherein the exogenous gene is an exogenous MinD gene.  
     
     
         31 . The vector according to  claim 30 , wherein said exogenous MinD gene is derived from  Arabidopsis thaliana  MinD gene.  
     
     
         32 . The vector according to  claim 28 , wherein the exogenous gene is an exogenous MinE gene.  
     
     
         33 . The vector according to  claim 32 , wherein said exogenous MinE gene is derived from  Arabidopsis thaliana  MinE gene.